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ERO1α promotes testosterone secretion in hCG‐stimulated mouse Leydig cells via activation of the PI3K/AKT/mTOR signaling pathway

PI3K/AKT/mTOR通路 间质细胞 细胞凋亡 蛋白激酶B 分泌物 睾酮(贴片) 免疫印迹 内科学 内分泌学 生物 细胞生长 细胞生物学 基因敲除 信号转导 化学 促黄体激素 医学 激素 生物化学 基因
作者
Fenglei Chen,Yujing Wang,Qinguang Liu,Jiahui Hu,Jiaqi Jin,Zhiyu Ma,Jinlong Zhang
出处
期刊:Journal of Cellular Physiology [Wiley]
卷期号:235 (7-8): 5666-5678 被引量:16
标识
DOI:10.1002/jcp.29498
摘要

Abstract ER oxidoreduclin 1α (ERO1α) is an oxidase, participating in formation of secretory and membrane proteins. However, the other physiological functions ERO1α is not well known. We found that ERO1α is high in the Leydig cells of the testis. Therefore, the purposes of the current study are to explore the role of ERO1α and the possible mechanisms in regulating cell proliferation, apoptosis, and testosterone secretion of Leydig cells. ERO1α was mainly localized in Leydig cells in the adult mice testes by immunofluorescence staining. Western blot analysis showed that ERO1α was higher in Leydig cells than that in the seminiferous tubules. The effect of ERO1α on cell proliferation, apoptosis, and testosterone secretion was detected by transducing ERO1α overexpression and knockdown lentiviruses into cultured primary Leydig cells (PLCs) together with hCG exposure. Flow cytometry analysis showed that ERO1α promoted cell proliferation by increasing cell distribution at the S phase and decreasing that at the G0/G1 phase. Western bolt analysis showed that ERO1α increased CDK2 and CDK6 expression. Cell apoptosis determination found that ERO1α inhibited PLC apoptosis. Western bolt analysis showed that ERO1α increased the ratio of BCL‐2/BAX, and decreased BAD and Caspase‐3 expression. Enzyme‐linked immunosorbent assay analysis demonstrated that ERO1α enhanced testosterone secretion. Western bolt analysis found that ERO1α increased StAR, 3β‐HSD, and CYP17A1 expression. Furthermore, ERO1α could activate the PI3K/AKT/mTOR signaling pathway. In summary, these results suggest that ERO1α might play proliferation promotion and antiapoptotic roles and enhance testosterone secretion in PLC, at least partly, via activation of the PI3K/AKT/mTOR signaling pathway.
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